Toc75-V/OEP80 is processed during translocation into chloroplasts, and the membrane-embedded form exposes its POTRA domain to the intermembrane space

FEBS Open Bio. 2020 Mar;10(3):444-454. doi: 10.1002/2211-5463.12791. Epub 2020 Feb 17.

Abstract

The insertion of membrane proteins requires proteinaceous complexes in the cytoplasm, the membrane, and the lumen of organelles. Most of the required complexes have been described, while the components for insertion of β-barrel-type proteins into the outer membrane of chloroplasts remain unknown. The same holds true for the signals required for the insertion of β-barrel-type proteins. At present, only the processing of Toc75-III, the β-barrel-type protein of the central chloroplast translocon with an atypical signal, has been explored in detail. However, it has been debated whether Toc75-V/ outer envelope protein 80 (OEP80), a second protein of the same family, contains a signal and undergoes processing. To substantiate the hypothesis that Toc75-V/OEP80 is processed as well, we reinvestigated the processing in a protoplast-based assay as well as in native membranes. Our results confirm the existence of a cleavable segment. By protease protection and pegylation, we observed intermembrane space localization of the soluble N-terminal domain. Thus, Toc75-V contains a cleavable N-terminal signal and exposes its polypeptide transport-associated domains to the intermembrane space of plastids, where it likely interacts with its substrates.

Keywords: OEP80; Toc75-V; chloroplasts; protein import; protein membrane insertion; β-barrel membrane protein.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Arabidopsis / genetics
  • Arabidopsis / metabolism
  • Arabidopsis Proteins / genetics
  • Arabidopsis Proteins / metabolism*
  • Chloroplasts / chemistry
  • Chloroplasts / genetics
  • Chloroplasts / metabolism*
  • Cytoplasm / metabolism
  • Intracellular Membranes / chemistry
  • Intracellular Membranes / metabolism
  • Membrane Proteins / genetics
  • Membrane Proteins / metabolism*
  • Membranes / metabolism
  • Mitochondria / metabolism
  • Nuclear Envelope / metabolism
  • Peas / genetics
  • Peas / metabolism
  • Protein Precursors / analysis
  • Protein Precursors / metabolism
  • Protein Transport

Substances

  • Arabidopsis Proteins
  • Membrane Proteins
  • OEP80 protein, Arabidopsis
  • Protein Precursors
  • TOC75 protein, Arabidopsis
  • Toc75-III protein, Arabidopsis